Characterization of extracellular circulating microRNA
Open accessAbstract
MicroRNAs (miRNAs), a class of post-transcriptional gene expression regulators, have recently been detected in human body fluids, including peripheral blood plasma as extracellular nuclease resistant entities. However, the origin and function of extracellular circulating miRNA remain essentially unknown. Here, we confirmed that circulating mature miRNA in contrast to mRNA or snRNA is strikingly stable in blood plasma and cell culture media. Furthermore, we found that most miRNA in plasma and cell culture media completely passed through 0.22 µm filters but remained in the supernatant after ultracentrifugation at 110 000g indicating the non-vesicular origin of the extracellular miRNA. Furthermore, western blot immunoassay revealed that extracellular miRNA ultrafiltrated together with the 96 kDa Ago2 protein, a part of RNA-induced silencing complex. Moreover, miRNAs in both blood plasma and cell culture media co-immunoprecipited with anti-Ago2 antibody in a detergent free environment. This is the first study to show that extracellular miRNAs are predominantly exosomes/microvesicles free and are associated with Ago proteins. We hypothesize that extracellular miRNAs are in the most part by-products of dead cells that remain in extracellular space due to the high stability of the Ago2 protein and Ago2-miRNA complex. Nevertheless, our data does not reject the possibility that some miRNAs can be associated with exosomes.
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Extracellular RNA is susceptible to degradation by circulating RNAses.
“Extracellular RNA is susceptible to degradation by circulating RNAses (2).”
From Analysis of Extracellular RNA by Digital PCR (2014) · cites this paper as [2]
The Results section (‘Extracellular versus intracellular miRNA expression pattern’; page unavailable) reports that synthetic RNA added to conditioned medium was immediately degraded, supporting susceptibility of unprotected extracellular RNA. But it also reports that endogenous extracellular miRNAs resisted added RNase A; the paper does not establish that extracellular RNA generally is degraded by circulating RNases. (LLM, conf=0.85)
Experimental — accuracy not yet measuredmiRNAs can be secreted into extracellular fluids and transported to target cells via vesicles such as exosomes or by binding to proteins including Argonautes.
“miRNAs can be secreted into extracellular fluids and transported to target cells via vesicles, such as exosomes, or by binding to proteins, including Argonautes.”
From Overview of MicroRNA Biogenesis, Mechanisms of Actions, and Circulation (2018) · cites this paper as [125]
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- Authors
- Turchinovich, A., Weiz, L., Langheinz, A., & Burwinkel, B.
- Affiliations
- German Cancer Research Center (DE)
- Corresponding author
- Turchinovich, Andrey
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 2011-05-24
- Volume
- 39
- Issue
- 16
- Pages
- 7223-7233
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- PMC
- PMC3167594
- OpenAlex
- W2167604259
- Cited by
- 1,941
- References
- 22
- Citations by year
- 2026: 59 · 2025: 65 · 2024: 76 · 2023: 94 · 2022: 111 · 2021: 181 · 2020: 156 · 2019: 150 · 2018: 152 · 2017: 160 · 2016: 145 · 2015: 184 · 2014: 147 · 2013: 142 · 2012: 98
- FWCI
- 44.16
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Nucleic Acids Research (publishedVersion, cc-by-nc)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc)
- PubMed Central (publishedVersion, cc-by-nc)
- PubMed (publishedVersion)
- Funders
- Deutsches Krebsforschungszentrum
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › MicroRNA in disease regulation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Keywords
- extracellular microRNAs, circulating microRNAs, blood plasma, Argonaute 2, RNA-induced silencing complex, microvesicles, exosomes, miRNA stability, post-transcriptional regulation
- MeSH terms
- Cell Line, Humans, Culture Media, Conditioned, RNA Stability, MicroRNAs, Exosomes, Argonaute Proteins
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